Hi, 

Using the TBARS assay, our OD readings are relatively stable.

If you send me your TBARS protocol, I could maybe offer some suggestions.

Anyway, I reccomend that you read all samples including the control at the same time (using a 96-well plate or other). 

Best,

Oren 

Are you making all measurements within 30min after stopping the reaction?

As Oren suggested, if possible, consider using a multiwell plate reader.

Dear Prem,

As Oren and Tausif reccomended, use a multiwell plate reader.

Dear all.

It is due to fact that your pooled organic solvent (butanol), wherein your all TBARS are remaining at the end of reaction, absorb humidity from the environment. So try to heat all of them for another 5-10 min after pooling the TBARS-containing layer, and take OD in a humidity-free room as quickly as possible.

This is the solution.

Good luck!!

Hi

as Oren said, our OD readings are  stable in TBARS. what is the procedure you do??

please check the following circumestances

1-heating the otienmixture reaction

2- cooling the mixture after heating

3-centrifuge the mixture to elimiate precipetated protien

Thanks every one for their valuable reply....I have uploaded the protocol I followed as an attachment to this question. Kindly have a look on it and please put forward some suggestions regarding the problem.